共刺激人外周血淋巴细胞对大肠癌细胞的杀伤作用

Experimental study of killed colon cancer cell by co-stimulated peripheral blood lymphocytes

目的探讨CD28单克隆抗体与CD80共刺激活化的人外周血淋巴细胞(PBLs)在体外对大肠癌细胞株HT-29杀伤强度及杀伤作用机制,为大肠癌的过继免疫治疗提供参考。方法获得PBLs；共刺激；检测活化细胞的体外淋巴细胞毒作用,并用电子显微镜观察效应细胞杀伤的大肠癌细胞超微结构,用流式细胞仪检测大肠癌细胞的凋亡指数。结果共刺激PBLs对肿瘤细胞杀伤作用较强,达到(69．35±1．17)%。电镜结果显示,效应细胞作用12h肿瘤细胞出现坏死,部分可见凋亡。流式细胞仪检测效应细胞凋亡指数为24 h 19．6%,48 h效应细胞凋亡指数为12．1%。结论活化的淋巴细胞杀伤大肠癌细胞是通过坏死及凋亡两条途径来实现的。

Objective To investigate the lethal effect of PBLs after monoclonal antibody CD28 ＋ CD80 co-stimulated on colon cancer cells in vitro. Adoptive immunotherapy is a way of possibility to cure colon cancer. Methods PBLs was obtained by Ficoll. The test groups were cultured with monoclonal antibody CD28 and CD80 co-stimulated in the experiment. The lymphocytes＇ cytotoxicity was examined by MTT methods. The microchange of the antipersonnel tumor cells was observed by electron microscope and the apoptotic index of colon cancer cells was examined by the flowcytometry. Results Colon cancer cells in vitro were killed by co-stimulated （69.35 ± 1.17） %. After 12 hours, cancer cells were observed necrosis and part of cells were found apoptosis through electron microscope. The index of the apoptosis was found to be 19.6% in 24 hours,12.1% in 48 hours through the examination of the flowcytometry. Conclusion Activated lymphocytes could kill the colon cancer cell with co-stimulated in two ways： necrosis and apoptosis.